利用蛋白质指纹图谱技术检测硒诱导金属硫蛋白实现大鼠肝纤维化防护作用  

Preliminary research on protein fingerprinting of metallothionein induced by selenium for the protection of liver fibrosis of rats

作　　者：雍政[1] 何冰[2] 徐卓立[3] 高燕[1] 董旭东[1] 黄英[1] 郭军华[1] 茹炳根[4] 

机构地区：[1]军事医学科学院附属医院临床药理室,北京100039 [2]武警医学院科研部,天津300162 [3]北京军医学院,北京100071 [4]北京大学生命科学院蛋白质工程国家重点实验室,北京100871

出　　处：《武警医学院学报》2007年第1期12-15,29,共5页Acta Academiae Medicinae CPAPF

基　　金：国家自然科学基金资助项目(30070884)

摘　　要：【目的】探索硒诱导金属硫蛋白及其对大鼠肝纤维化防护作用的蛋白质指纹图谱改变。【方法】利用表面增强的激光解析离子化-飞行时间-质谱(SELDI-TOF-MS)技术测定硒对金属硫蛋白的诱导,寻找大鼠肝纤维化的异常蛋白表达标志并评估硒代金属硫蛋白的防护作用。【结果】IMAC-Cu蛋白质芯片6279 Da和6650Da处特征性MT蛋白质表达增强证实硒对MT的高诱导作用;肝组织点样的WCX2芯片5 140Da综合峰的两峰与正常组相对应处比较明显倒置,血清点样的H4芯片7 105 Da左右的蛋白质出现高表达,4 820Da左右的蛋白质表达受抑,三种图谱变化是大鼠酒精性肝纤维化的相关信号蛋白;硒诱导MT增高后上述异常表达均有改善,说明有机硒对肝纤维化有防护作用。【结论】组织和血清的蛋白质指纹图谱可以作为肝纤维化疗效监测和评价的一类标志物,为今后进行更多的药物或方法筛选奠定了一种可以尝试的试验模式。[Objective] To explore the protein fingerprinting of metaUothionein （MT） induced by selenium to protect the rat liver from fibrosis. [Methods] The effects of selenium to induced MT in liver of rats were determined by surface enhanced laser desorption ionization-time of flight mass spectrometry （SELDI-TOF-MS）o Finding the unnormal protein-labels of liver fibrosis in tissue and blood serum of rats and the preventive effects of Se-MT against the liver fibrosis of rats were evaluated. [Results] Through SELDI-TOF-MS the protein fingerprinting of liver tissue on IMAC-Cu chip expressed the peak current intensity in 6 279 Da and 6 650 Da increased. The ratio of peak current intensity in 5 130 Da / 5 142 Da was upside down in the injury group, but not in control and Se pre-treated groups on WCX2 chip. On H4 chip the expression of peak current intensity in 7 105 Da was increased and that in 4 820 Da was decreased. [Conclusion] This finding gives a good reference protein label and proves that Se-MT with protective effects against liver fibrosis. The protein fingerprinting may provide a novel biological signs and experimental model for the diagnosis and therapeutic evaluation about liver fibrosis.

关 键 词：硒 金属硫蛋白 蛋白质指纹图谱/SELDI—TOF—MS 肝纤维化 

分 类 号：R575.2[医药卫生—消化系统]